Product and process for transformation of thraustochytriales microorganisms

Abstract

Disclosed are nucleic acid and amino acid sequences for acetolactate synthase, acetolactate synthase regulatory regions, α-tubulin promoter, a promoter from a Thraustochytriales polyketide synthase (PKS) system, and fatty acid desaturase promoter, each from a Thraustochytriales microorganism. Also disclosed are recombinant vectors useful for transformation of Thraustochytriales microorganisms, as well as a method of transformation of Thraustochytriales microorganisms. The recombinant nucleic acid molecules of the present invention can be used for the expression of foreign nucleic acids in a Thraustochytriales microorganism as well as for the deletion, mutation, or inactivation of genes in Thraustochytriales microorganisms.

The invention claimed is: 
     
       1. A method for production of a protein, comprising:
 culturing a recombinant microorganism of the Order Thraustochytriales in a medium, wherein the recombinant microorganism comprises an isolated nucleic acid molecule comprising a foreign gene that encodes the protein, to produce the protein; and recovering the protein. 
 
     
     
       2. The method of  claim 1 , wherein the isolated nucleic acid molecule further comprises one or more transcription control sequences and the foreign gene is operatively linked to the one or more transcription control sequences. 
     
     
       3. The method of  claim 2 , wherein the one or more transcription control sequences comprise a Thraustochytriales transcription control sequence. 
     
     
       4. The method of  claim 3 , wherein the Thraustochytriales transcription control sequence is selected from the group consisting of Thraustochytriales promoter sequences, Thraustochytriales enhancer sequences, Thraustochytriales operator sequences, Thraustochytriales repressor sequences, and combinations thereof. 
     
     
       5. The method of  claim 4 , wherein at least one Thraustochytriales transcription control sequence is a Thraustochytriales promoter sequence. 
     
     
       6. The method of  claim 5 , wherein the Thraustochytriales promoter sequence is selected from the group consisting of a Thraustochytriales acetolactate synthase promoter sequence, Thraustochytriales α-tubulin promoter sequence, Thraustochytriales polyketide synthase system promoter sequence, and Thraustochytriales fatty acid desaturase promoter sequence. 
     
     
       7. The method of  claim 1 , wherein the isolated nucleic acid molecule further comprises a signal segment nucleic acid sequence. 
     
     
       8. The method of  claim 1 , wherein the protein is secreted into the medium, and wherein the protein is recovered from the medium. 
     
     
       9. The method of  claim 1 , wherein the microorganism is from a genus selected from the group consisting of  Thraustochytrium, Labyrinthuloides, Japonochytrium , and  Schizochytrium.    
     
     
       10. The method of  claim 9 , wherein the microorganism is from a species selected from the group consisting of  Schizochytrium  sp.,  Schizochytrium aggregatum, Schizochytrium limacinum, Schizochytrium minutum, Thraustochytrium  sp.,  Thraustochytrium striatum, Thraustochytrium aureum, Thraustochytrium roseum , and  Japonochytrium  sp. 
     
     
       11. A method for production of a protein, comprising:
 culturing a recombinant microorganism of the Order Thraustochytriales in a medium, wherein the recombinant microorganism comprises an isolated nucleic acid molecule comprising a foreign gene that encodes the protein, to produce the protein; and harvesting the microorganism comprising the protein. 
 
     
     
       12. The method of  claim 11 , wherein the microorganism is harvested as a biomass.